人参皂苷Rg-1对人内皮祖细胞增殖、凋亡和分泌HGF的影响

Effects of Ginsenoside Rg-1 on Proliferation, Apoptosis and HGF Secretion of Human Endothelial Progenitor Cells

目的 探讨人参皂苷Rg-1(ginsenoside Rg1,GS-Rg1)对人内皮祖细胞(endothelial progenitor cells, EPCs)体外增殖和分泌肝细胞生长因子(hepatocyte growth factor, HGF)的影响,为进一步研究GS-Rg1通过EPCs介导血管新生的作用机制奠定基础。方法 取正常剖宫产胎盘中的脐带血,密度梯度离心分离出单个细胞;通过CD31免疫荧光鉴定和双荧光染色的共聚焦拍摄(DiI-acLDL和FITC-UEA-1配合DAPI染核)共同对EPCs进行鉴定;将鉴定成功的EPCs分别在不同浓度梯度Gs-Rg1(0、5、10、2、40、80 mg/L)下干预培养48 h,确定Gs-Rg1促EPCs增殖的最佳浓度。另将获得的EPCs随机分为实验组(最佳浓度Gs-Rg1干预)和对照组(PBS干预)。采用细胞计数试剂盒(Cell Counting Kit-8,CCK-8)、流式细胞术及ELISA法分别检测GS-Rg1对EPCs的增殖、凋亡和分泌HGF的影响。结果 在一定范围(<40 mg/L)内,GS-Rg1的浓度与EPCs增殖呈正相关,Gs-Rg1促EPCs增殖的最佳浓度为40 mg/L;与对照组相比,实验组细胞增殖、抗凋亡和分泌HGF的能力显著增强,差异显著,具有统计学意义(P<0.05)。结论 GS-Rg1对EPCs无毒性,且能改善其增殖功能和促进HGF的分泌。

Objective It aims to study the effect of ginsenoside Rg-1 on proliferation and secretion of hepatocyte growth factor(HGF)of human endothelial progenitor cells in vitro and provide the basis for the further study on the mechanism of GS-Rg1 mediating angiogenesis through EPCs.Methods The umbilical cord blood from placenta of normal cesarean section was obtained and single cells were separated by density gradient centrifugation.EPCs were identified by CD31 immunofluorescence identification and confocal shooting with double fluorescence staining(DiI-acLDL and FITC-UEA-1 combined with DAPI staining).The successfully identified EPCs were cultured under different concentration gradients Gs-Rg1(0,5,10,20,40,80 mg/L)for 48 hours to determine the best concentration of Gs-Rg1 to promote EPCs proliferation.The EPCs were randomly divided into experimental group(intervention with the best concentration of Gs-Rg1)and control group(intervention with PBS).The effects of GS-Rg1 on proliferation, apoptosis and HGF secretion of EPCs were detected by cell counting kit-8,flow cytometry and ELISA.Results There was a positive correlation between GS-Rg1 concentration and EPCs proliferation in a certain range(<40 mg/L).The optimal concentration of Gs-Rg1 to promote EPCs proliferation was 40 mg/L.Compared with those of the control group, the ability of cell proliferation, anti-apoptosis and HGF secretion in the experimental group were significantly enhanced(P<0.05).Conclusion GS-Rg1 has no toxicity to EPCs and improves its proliferation function and promotes HGF secretion.